Isolation and identification of urinary nucleosides. Applications of high-performance liquid chromatographic methods to the synthesis of 5'-deoxyxanthosine and the simultaneous determination of 5,6-dihydrouridine and pseudouridine.
Modified nucleosides from pooled normal human urine were extracted using a boronate affinity gel column and fractionated by reversed-phase high-performance liquid chromatography (RP-HPLC). The major constituents in each of the 30 RP-HPLC fractions were determined by gas chromatography-mass spectrometry of the trimethylsilyl derivatives of the fractions. The same RP-HPLC method was used in the synthesis of 5'-deoxyxanthosine from authentic 5'-deoxyadenosine. In addition, the simultaneous determination of urinary 5,6-dihydrouridine (D) and pseudouridine (psi) was carried out by RP-HPLC using two ODS columns in series. The level of D in pooled normal urine was 4.87 nmol/mumols creatinine. The RP-HPLC method was applied to the measurement of D and psi levels in urines collected before and after surgery from four patients with gastrointestinal cancer. A large decline in both nucleoside levels in urines after surgery was observed in three of the four cancer patients.
Nakano
K
K
Perfect Liberty Comprehensive Research Institute, Osaka, Japan.
Yasaka
T
T
Schram
K H
KH
Reimer
M L
ML
McClure
T D
TD
Nakao
T
T
Yamamoto
H
H
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003401:Creatine / Q000652:urine; D003853:Deoxyribonucleosides; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D005770:Gastrointestinal Neoplasms / Q000652:urine; D006801:Humans; D008297:Male; D009705:Nucleosides / Q000302:isolation & purification / Q000652:urine*; D011560:Pseudouridine / Q000652:urine; D012263:Ribonucleosides / Q000096:biosynthesis; D014529:Uridine / Q000031:analogs & derivatives / Q000652:urine
D003853:Deoxyribonucleosides; D009705:Nucleosides; D012263:Ribonucleosides; C067231:5'-deoxyxanthosine; D011560:Pseudouridine; C010922:5,6-dihydrouridine; D003401:Creatine; D014529:Uridine